Neural stem cells (The Johns Hopkins Medical Institutions)

Clear spinal cord tissue of meninges and dissociate dorsal root ganglia into a single-cell suspension by mechanical dissociation in serum-free, modified N2 medium. Serially expand in monolayer. Change growth medium every other day, and, on alternate days, add basic fibroblast growth factor (bFGF; 10 ng/ml) to the culture. Conduct the first passage 16 days postplating, a time point at which the culture is mostly composed of dividing NSCs and postmitotic neurons.